D'Ambrosio began his career in karting, between 1995 and 2002. He was three times Belgian champion, winning the Mini class in 1996, Junior in 2000 and Formula A in 2002. He won the Junior Monaco Kart Cup in 2000, and won World Cup Formula A in 2002.

D'Ambrosio moved into single-seaters in 2003, and won the Belgian Formula Renault (1.6 litre) championship. He also drove in the German-based Formula König championship in 2003. He moved into the French Formula Renault (2-litre) series for 2004. He finished fourth in the series and was top rookie. He switched to the Italian Formula Renault Championship in 2005, and finished fourth. In 2006, he drove in the Formula Renault 3.5 Series, for Tech 1 Racing. He left the series after four rounds and zero points.

During this time, d'Ambrosio also raced in several Eurocup races. In 2006, he raced in the second half of the Euroseries 3000. D'Ambrosio had three podium finishes and finished fifth in the point standings.

For 2008, d'Ambrosio raced in both the GP2 Series and the new GP2 Asia Series. He finished eleventh in the GP2 drivers' championship. In the 2008-09 GP2 Asia Series season, d'Ambrosio finished second to team-mate Kamui Kobayashi. In the 2009 season, he finished ninth with out wining a single race. In 2010, d'Ambrosio took his first series victory at Monaco. He took his first GP2 pole position at Belgian. He finished twelfth in the drivers' championship.

In January 2010, d'Ambrosio was announced as a reserve driver for Renault F1. He replace Lucas di Grassi at Virgin four times for Friday practice: at the Singapore, Japanese, Korean and Brazilian Grands Prix.

On 21 December 2010 Virgin Racing confirmed that d'Ambrosio would race for the team in 2011.[1]